HAECs were grown on 8-well slides, incubated with flavoring compounds for 90 minutes, fixed with 2% paraformaldehyde, and stored at −80°C. A commercially available TUNEL assay (terminal deoxynucleotidyl transferase dUTP nick-end labeling; Roche) was performed, and cells were imaged on a fluorescence microscope (Nikon Eclipse TE2000) for fluorescein and DAPI (4’,6-diamidino-2-phenylindole; Vector Laboratories). DNAse 1 (Sigma) was used as a positive control for apoptosis. A minimum of 50 cells were quantitated for each condition. Data are presented as % TUNEL-positive cells.
Terminal deoxynucleotidyl transferase dutp nick end labeling
Manufactured by Roche
Sourced in China, Switzerland
Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) is a method used to detect and quantify apoptosis (programmed cell death) in cells. It is a technique that labels the terminal end of nucleic acid fragments generated during apoptosis.
Automatically generated - may contain errors
3 protocols using terminal deoxynucleotidyl transferase dutp nick end labeling
1
Quantifying Apoptosis in HAECs
2
Testicular and Pituitary Histology
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The pathology of the testes and pituitaries in hematoxylin–eosin, Masson trichome (JianCheng Bioengineering Institute, Nanjing, China) and terminal deoxynucleotidyl transferase dUTP nick‐end labeling (Roche, Basel, Switzerland) stained sections was examined by light microscopy. The testes and pituitary glands were dissected into small cubes (1 mm3), fixed in 4% pre‐cooling glutaraldehyde and then fixed using osmium acid, dehydrated with an acetone gradient, soaked in epoxy resin, embedded, sliced, and stained with uranium acetate and citric acid lead.
The ultrastructure of the testes and pituitary glands was examined with an H‐7500 transmission electron microscope (Hitachi, Tokyo, Japan).
The ultrastructure of the testes and pituitary glands was examined with an H‐7500 transmission electron microscope (Hitachi, Tokyo, Japan).
3
Apoptosis Quantification by TUNEL
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Terminal deoxynucleotidyl transferase dUTP nick-end labeling was performed according to kit manufacturer instructions (Roche, Basel, Switzerland) . Five fields at 400X magnification were randomly selected from each section; the average number of apoptotic cells in 1000 cells was calculated and reported as percentage (apoptosis index: AI).
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